不同固定条件对几种植物样品超微结构的影响

The Improvement of the Sample Preparation Technique for TEM of Plant

为比较几种固定条件对不同植物材料样品电镜超微结构图像清晰度的影响。以3个不同科属4种植物[油棕(Elaeis gineansis Jacq.)、拟南芥(Arabidopsis thaliana.)、花生(Arachis hypogaea Linn.)、柱花草(Stylosanthes guianensias Sw.)]为研究主体代表,探讨了2.5%戊二醛与4%戊二醛固定过夜、锇酸固定2 h及过夜的制备方法对其叶片超微结构的影响,并成功观察了柱花草接种胶苞炭疽野生型菌株CH008后的病菌侵入过程。结果表明:油棕、拟南芥、花生及柱花草叶片经4%戊二醛与1%锇酸固定液固定过夜,其组织成像的效果均较为理想,各种细胞器整体信息丰富、结构组织完好、线条清晰。表明,此制备方法能够较好的保存植物叶片和细胞组织,提高制样的成功率及观察分辨率。

In this study, we analyzed the sample preparation from three different species for verifying available methods and application. For ultrastructural effect of Elaeis guineensis, Arabidopsis thaliana, Arachis hypogaea,Stylosanthes guianensias, we compared some methods, i.e. 2.5% of glutaraldehyde, 4% of glutaraldehyde, osmic acid（fixed for 2 h and overnight）, and the infection process of wide-type strain CH008 in S. guianensias inoculated with Colletotrichum gloeosporioides was also observed. The results showed that there was no significant difference on the effect of ultramicro images of organelle when the tissues were fixed with 4% of glutaraldehyde and 2.5%of glutaraldehyde for A. thaliana, A. hypogaea and S. guianensias. However, for E. guineensis, the image effect of tissues treated with 4% of glutaraldehyde was better compared to the treatment of 2.5% of glutaraldehyde, and the effect of the leaves fixed for overnight was better than that for 2 h. Therefore, the leaf tissues would not get more injuries by the method（ samples were fixed with 4% of glutaraldehyde and 1% osmic acid for overnight）, which meaning that the method wouldl improve resolution ratio and be helpful for phytopathology studies.